Expression of Tmem119/Sall1 and Ccr2/CD69 in FACS-Sorted Microglia- and Monocyte/Macrophage-Enriched Cell Populations After Intracerebral Hemorrhage
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Activation and polarization of microglia and macrophages are critical events in neuroinflammation and hematoma resolution after intracerebral hemorrhage (ICH). However, distinguishing microglia and monocyte-derived macrophages histologically can be difficult. Although they share most cell surface markers, evidence indicates that the gene regulation and function of these two cell types might be different. Flow cytometry is the gold standard for discriminating between the two cell populations, but it is rarely used in the ICH research field. We developed a flow cytometry protocol to identify and sort microglia and monocyte-derived macrophages from mice that have undergone well-established ICH models induced by collagenase or blood injection. In addition, we combined a recently established magnetic-activated cell separation system that allows eight tissue samples to be assessed together. This protocol can be completed within 5-8 h. Sorted cells are fully preserved and maintain expression of microglia-specific (Tmem119/Sall1) and macrophage-specific (Ccr2/CD69) markers. They retain phagocytic ability, respond to lipopolysaccharide stimulation, and engulf fluorescent latex beads. Thus, this protocol represents a very important tool for researching microglial and monocyte-derived macrophage biologic function after ICH and other brain diseases.
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Analyses using CD45- and CD11b-specific antibodies have been used to effectively identify microglial and macrophage populations in the normal retina.
“Analyses using CD45- and CD11b-specific antibodies have been used to effectively identify microglial and macrophage populations in the normal retina.17, 18, 19”
From Characterizing the cellular immune response to subretinal AAV gene therapy in the murine retina (2021) · cites this paper as [18]
The Results (Flow Cytometry and FACS) report that CD11b and CD45 staining clearly separated microglia from monocytes/macrophages in mouse brains 24 hours after intracerebral hemorrhage. The paper does not establish that these antibodies effectively identify those populations in the normal retina; its sham samples were also brain samples. (LLM, conf=0.88)
Experimental — accuracy not yet measuredCo-expression of CD45 and CD11b has effectively been used to distinguish microglia and macrophages.
“However, although the co-expression of CD45 and CD11b has effectively been used to distinguish microglia and macrophages,17, 18, 19, 20,49 microglia can increase CD45 expression following activation, which may limit the resolution between these two populations in the inflamed retina.50,51”
From Characterizing the cellular immune response to subretinal AAV gene therapy in the murine retina (2021) · cites this paper as [18]
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Self-citation
- Authors
- Li, Q., Lan, X., Han, X., & Wang, J.
- Affiliations
- Johns Hopkins University (US)
- Corresponding author
- Wang, Jian
- Venue
- Frontiers in Cellular Neuroscience
- Publisher
- Frontiers Media SA
- Published
- 2019-01-09
- Volume
- 12
- Pages
- 520-520
- ISSN
- 1662-5102 (ISSN-L)
- PMC
- PMC6333739
- OpenAlex
- W2909345676
- Cited by
- 64
- References
- 62
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 10 · 2023: 14 · 2022: 8 · 2021: 13 · 2020: 9 · 2019: 6
- FWCI
- 3.96
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Frontiers in Cellular Neuroscience (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Institutes of Health (R01NS078026, R01AT007317, R56NS096549)
- American Heart Association (17GRNT33660766, 16POST29640010, 17POST3366019, 16SDG30980031)
- Topics
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Health Sciences › Medicine › Neurology › Intracerebral and Subarachnoid Hemorrhage Research
- Health Sciences › Medicine › Neurology › Traumatic Brain Injury and Neurovascular Disturbances
- Keywords
- intracerebral hemorrhage, microglia, monocyte-derived macrophages, FACS cell sorting, hematoma resolution, neuroinflammation, phagocytosis, collagenase-induced hemorrhage
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